Cellooligosaccharide is a generic name for cellobiose, cellotriose, cellotetraose, cellopentaose, and cellohexaose and is an oligosaccharide with 1 to 6 glucopyranose units linked by a β-1,4 bond in one molecule.
In the present specification, a composition containing other components, for example, glucose in addition to these cellooligosaccharides is referred to as a “cellooligosaccharide composition.”
Furthermore, in the present specification, a composition that comprises this “cellooligosaccharide composition” as an active ingredient and may contain other additives or the like is referred to as a “cellooligosaccharide-containing composition” or “drug composition”).
Cellooligosaccharide are hardly digested and absorbed by humans, and therefore, they are useful as low-calorie sweeteners. In addition, they are also useful as general foods, functional foods, cosmetics, drugs and additives hereof, other raw materials for chemical conversion, and raw materials for fermentation (Non-patent Document 1).
With a focus on lipids in the liver that can cause pathological conditions such as diabetes, arteriosclerosis, and cirrhosis in adults, the present invention can be used as an agent for preventing or improving a lifestyle-related disease because a composition comprising a cellooligosaccharide composition with a specific saccharide composition suppresses a decrease of a blood adiponectin concentration and decreases lipids in the liver.
Non-patent Document 2 discloses bioavailability of dietary intake calorie, blood sugar levels, blood insulin concentrations, and the like when healthy humans (females) ingest a cellooligosaccharide composition consisting of 92.9% of cellobiose, 3.3% of cellooligosaccharides with a degree of polymerization of 3 or higher, and 3.8% of glucose. In the document, only blood sugar levels and insulin concentrations were measured to confirm digestibility of the cellooligosaccharide composition, and the composition was only orally administered to humans to obtain exact dietary intake calories. Furthermore, the document includes no description or suggestion about the intake of the cellooligosaccharide composition and the neutral fat concentration in the liver. Therefore, the document is completely different from the cellooligosaccharide composition of the present invention, which has a specific saccharide composition and a low decreasing rate of blood adiponectin, and is used as an agent for preventing or improving a lifestyle-related disease since the composition is effective in decreasing the neutral fat concentration in the liver by oral intake thereof.
The following reports are available on the attempts to use cellooligosaccharides for lipid metabolization.
Non-patent Documents 1, 3, and 4 describe results after breeding SD male rats for 4 weeks by feeding a high sucrose diet (64.7 parts by mass of sucrose, 25 parts by mass of casein, 5 parts by mass of corn oil, 4 parts by mass of mineral, 1 part by mass of vitamins, 0.2 parts by mass of choline hydrochloride, and 0.05 parts by mass of vitamin E) of which 1 or 2.5% by mass was replaced with a cellooligosaccharide composition comprising 85.7% by mass of cellobiose, 3.7% by mass of cellotriose, and 9.3% by mass of glucose as a saccharide composition. According to the document, intake of the cellooligosaccharide composition having the above-mentioned saccharide composition decreased serum total cholesterol, HDL-cholesterol, and triglyceride levels and body fat rates as compared with a group without the cellooligosaccharide composition. However, although the cellooligosaccharide of the documents showed a decreasing effect of serum lipid, the weights of the internal organs such as a liver did not change. Furthermore, a decreasing effect of visceral lipid was not observed. On the other hand, the present invention provides an agent for preventing or improving a lifestyle-related disease that has a specific saccharide composition; suppresses a decrease of a blood adiponectin concentration; comprises a cellooligosaccharide composition; and has an excellent decreasing effect of lipid in the liver. Therefore, it is totally different from the cellooligosaccharide of the documents decreasing the serum lipid.
Furthermore, Patent Document 1 describes food/feed additives for regulating intestinal functions and promoting lipid metabolism which are characterized in that lactic bacteria cells (Lactobacillus rhamnosus) and cellobiose are contained in mixture ratios of 1:2.5 to 8. The document describes results after breeding male Wister rats for 14 days by feeding an elemental diet 20.0 parts by mass of casein, 0.3 parts by mass of DL-methionine, 3.5 parts by mass of minerals, 1.0 parts by mass of vitamins, 7.0 parts by mass of soybean oil) of which 10% by mass was replaced with cellobiose. According to the document, the group fed with diet containing cellobiose alone had no effect on serum or visceral lipid metabolism as compared with the group fed with diet not containing the same. The lipid levels were decreased only when cellobiose and the above-mentioned lactic bacterium were used in combination. Therefore, it is essentially different from an agent for preventing or improving a lifestyle-related disease such as the saccharide composition of the present invention, which comprises cellooligosaccharides having a highly regulated saccharide composition, and decreases fat levels in the liver by cellooligosaccharides alone. Furthermore, since the diet for promoting the lipid metabolism of the document requires simultaneous intake of cellobiose and viable bacterial cells, there is a problem that the effect thereof decreases to a considerable extent before use because of poor stability of viable bacterial cells in a formulation.
Therefore, there has been known no lifestyle-related disease preventing or improving agent which comprises, as an active ingredient, an Insulin-non-increasing cellooligosaccharide composition having a specific saccharide composition and suppressing a decrease in a blood adiponectin concentration and decreases lipids in the liver by taking the same.
Hereafter, activation of the enteric bacteria flora by the cellooligosaccharide composition of the present inanition or the cellooligosaccharide-containing composition of the present invention will be explained. In recent years, it has been revealed that the enteric bacteria flora is closely related to human health. For example, bifidobacteria, lactic bacteria and the like are useful bacteria that bring health to humans. Since they decrease with aging, attempts have been made to improve the enteric bacteria flora (intestinal flora) with functional foods such as prebiotics (served as nutrients for enteric bacteria and having a function to increase them). There are many existing functional foods that propagate bifidobacteria and lactic bacteria.
In particular, the above-mentioned prebiotics with a higher selective activity of activating only useful intestinal bacteria and not activating harmful bacteria are more efficient. Examples of the existing attempts include foods that activate useful bacteria while inhibiting propagation of Clostridium perfringens, a kind of a Welch bacillus, as harmful intestinal bacteria.
Among the above-described attempts of increasing bifidobacteria and lactic bacteria and suppressing Wench bacillus, those related to oligosaccharides with β bonds will be described below.
Patent Document 2 describes the substance for improving intestinal flora comprising glucoolroosaccharides having β-glucosidic bonds and/or a reduced product thereof as an active ingredient. The glucooligosaccharide described in the document is one or two or more types selected from cellobiose, sophorose, laminaribiose, gentiobiose, and gentiooligosyl-D-glucose. This oligosaccharide undoubtedly promotes propagation of useful bacteria such as bifidobacteria and lactic bacteria and has an effect of suppressing propagation of Welch bacillus. 
However, there was a problem that harmful bacteria other than Welch bacillus such as Bacteroides fragilis and Eubacterium aerofaciens were increased to a considerable extent. Bacteroides fragilis and Eubacterium aerofaciens are anaerobic asporogenic Gram-negative bacillus which are harmful bacteria relating to various abscess forming infections and bacteremia caused by the mixed infection thereof with aerobic bacteria. The agent for improving intestinal flora of the above-mentioned document is totally different from the present invention of an agent for activating enteric bacteria which can propagate useful bacteria such as bifidobacteria and lactic bacteria and suppressing propagation of harmful bacteria such as Bacteroides fragrlis and Eubacterium aerofaciens. Therefore, there has been known no agent for activating enteric bacteria that has a regulated saccharide composition of cellooligosaccharides in a specific range; propagates useful bacteria such as bifidobacteria and lactic bacteria; and suppresses propagation of harmful bacteria such as Bacteroides fragailis and Eubacterium aerofaciens. 
Hereafter, suppression of propagation of Helicobacter pylori or a bacteriostatic effect against Helicobacter pylori by the cellooligosaccharide composition or the cellooligosaccharide-containing composition of the present invention will be described. Helicobacter pylori is a microaerobic Gram-negative bacillus. Helicobacter pylori is thought to live in a strongly acidic environment in the stomach by producing ammonia from urea by the urease activity thereof to neutralize the gastric acid. It has been revealed that this ammonia produced by Helicobacter pylori damages the stomach mucous membrane and it is associated with stomach diseases such as gastritis, ulcer, stomach cancer, and lymphoma. It is recognized that suppression or regulation of the growth of Helicobacter pylori is an effective method for prevention or improvement of these diseases.
As a conventional method for suppressing Helicobacter pylori, administration of antibiotics has been attempted. This method undoubtedly has an effect of suppressing Helicobacter pylori. However, there was also a problem that the suppressing actions of the antibiotics affected not only Helicobacter pylori, but also enteric bacteria which are useful for the human body. Furthermore, there was also a problem that the antibiotics had no small effects on the human body such as causing diarrhea as the adverse drug reaction. Therefore, a suppressing agent that is safe to the human body and acts selectively on Helicobacter pylori has been desired. As prior art, examples of the agent for suppressing Helicobacter pylori comprising a saccharide as an active ingredient include the following.
Patent Document 3 discloses an antitumor agent comprising fucoidan as an active ingredient. The antitumor agent of this document utilizes inhibition of Helicobacter pylori colonization on the stomach wall by fucoidan and has effects of preventing infection of Helicobacter pylori. On the other hand, since the agent for suppressing Helicobacter pylori of the present invention comprising a cellooligosaccharide as an active ingredient directly suppresses Helicobacter pylori irrespective of colonization on the stomach wall, it has an effect of improvement in addition to prevention and is totally different from the preventing-agent of the document.
The present invention is a composition comprising a cellooligosaccharide composition with a specific saccharide composition as an active ingredient. There has been known no agent for suppressing Helicobacter pylori or no bacteriostatic agent against Helicobacter pylori that is safe to the human body which suppresses propagation of Helicobacter pylori by orally taking.
Hereafter, the property of the cellooligosaccharide composition for improving the skin barrier function or the cellooligosaccharide-containing composition of the present invention will be described. In the human skin, the horny cell layer functions as a barrier against the external environment of an organism. The barrier function of the skin is closely related to the amount of transepidermal water loss of the skin, and suppression of this transepidermal water loss is critical in maintaining flexibility and moisture of the skin (Non-patent Documents 5 and 6). So far, inventions have been made with a focus on regeneration of the lamellar structure of the horny cell layer using saccharides or oligosaccharides as an active ingredient.
Patent Document 4 describes an agent regenerating the lamellar structure of the horny cell layer cells in the skin that comprises glucose and/or raffinose, and an agent regenerating the lamellar structure of the horny cell layer cells in the skin characterized in that glucose and oligosaccharides are contained.
The document describes, as a lamellar structure regenerating agent, a topical agent comprising glucose as an essential component and one or more oligosaccharides selected from the group consisting of raffinose, melibiose, trehalose, sucrose, maltose, cellobiose, gentianose, stachyose, and cyclodextrin. According to this document, a lamellar structure regenerating effect is undoubtedly observed in raffinose, melibiose, trehalose, sucrose, and cyclodextrin, and an effect of decreasing “greasiness” is further observed by combination use of glucose and raffinose. However, this document has no description about recovery of the transepidermal water loss by the above-mentioned cellooligosaccharides including cellobiose. In particular, it is not known that cellooligosaccharides are excellent in usability and promotion of recovery of the transepidermal water loss and improve the skin barrier function without addition of glucose. Furthermore, since the agent for improving lamellar structure of this document needs to contain a large amount of glucose such as 7.4% by mole or more in proportion to raffinose or oligosaccharides, there was a problem that, when it was used in combination with an amino acid or a component containing an amino group such as a protein and subjected to a heating process, browning or discoloration occurred due to the Maillard reaction, which markedly reduced the commodity value. The present invention is essentially different from the above-described invention in that it has minimal “greasiness”; decreases browning; and can improve the skin barrier function without containing glucose as an essential component. Therefore, there has been known no agent for improving skin barrier function of the present invention that comprises cellooligosaccharides including cellobiose in a specific range as an active ingredient; has favorable usability; causes minimal browning; and promotes recovery of transepidermal water loss without using glucose in combination.
Hereafter, the Indigenous dermal bacteria flora improving property of the cellooligosaccharide or the cellooligosaccharide composition of the present invention will be described.
The healthy human skin maintains epidermal pH as weakly acidic and is protected by beneficial indigenous dermal bacteria having an action for improving resistance. However, internal tissues of individuals with easily damaged skin due to dry skin, atopic dermatitis, or the like are infected with Staphylococcus aureus and Pseudomonas aerugunosa, harmful bacteria, from the injury on the skin surface. The infection causes further deterioration of the human skin condition. When internal tissue is exposed, Staphylococus aureus and Pseudomonas aeruginosa are attached to the injury and grow on the exudated body fluid even if the injury is as small as invisible. In fact, it is said that a large amount off Staphylococcus aureus is detected in the skin of patients with atopic dermatitis. Therefore, it is extremely important to maintain the cell count of beneficial dermal bacteria and suppress the cell count of Staphylococcus aureus and Pseudomonas aeruginosa, harmful bacteria on the human skin.
Furthermore, among the skins, more indigenous dermal bacteria exist in the skin of the scalp where hairs are more densely grown than other sites, and the balance of the cell counts of these bacteria is very important. Furthermore, it is also important to impart refreshing feeling and smoothness to dry hair as sensory effects.
Many drugs and raw material components for cosmetics having a bacteriostatic action against Staphylococcus aureus or Pseudomonas aeruginosa have been disclosed. Examples thereof include N-acylglutamic acid-containing detergents (Patent Document 5, iron-bound lactoferrin-containing agents Patent Document 6), various plant extracts (Patent Documents at 8, 9, 10, and 11), and so forth. However, since these agents also have a bacteriostatic action against beneficial indigenous dermal bacteria with potency comparable to or greater than the action against bacteria belonging to genus Staphylococcus, they could not be used as agents for improving indigenous dermal bacteria flora.
Furthermore, an agent for adjusting the balance of the counts of Living indigenous dermal bacteria by mixing a bittern and a rosemary extract or a licorice extract has been disclosed (Patent Document 12). However, the effect was not adequate, and bacteriostasis against Pseudomonas aeruginosa was not described. Furthermore, refreshing feeling or smoothness was not imparted as a touch in a dried condition, and the agent was expensive and was very difficult to use in practice.
Furthermore, an agent for improving indigenous dermal bacteria flora comprising isomaltooligosaccharide has been disclosed (Patent Document 13). According to this document, an agent for improving indigenous dermal bacteria flora comprising isomaltooligosaccharide and/or a sugar alcohol obtained by reducing isomaltooligosaccharide does not act on staphylococcus ecidermidis in a specific range of pH and amount thereof and has an effect of suppressing Staphylococcus aureus. As shown in the comparative examples of the present application, however, these effects are not adequate when high amounts of carbohydrate a added in order to improve the retention of skin moisture and a touch of a face lotion. Furthermore, the document has no description about bacteriostasis against Pseudomonas aeruginosa, and the agent did not impart refreshingness or smoothness as a touch in a dried condition.
Therefore, the agent for improving dermal bacteria flora of the document is fundamentally different from the agent for improving indigenous dermal bacteria flora of the present invention, which does not exhibit a bacteriostatic action against a beneficial dermal bacterium, i.e. Staphylococcus epidermidis; but exhibits a bacteriostatic action or a propagation-suppressing action against harmful bacteria, i.e. Staphylococcus aureus and Pseudomonas aeruginosa; and imparts refreshingness or smoothness as a touch of dried hair.
Finally, the high fluidity cellooligosaccharide powder of the present invention which is excellent in oil retention in addition to powder fluidity and uniform dispensability; hardly absorbs moisture; and is excellent in handling properties such as a compression moldability, heat resistance, acid resistance, a property of preventing starch retrogradation, and a property of preventing protein modification will be described below.
Patent Document 14 describes a solid formulation comprising cellobiose as the principal ingredient and cellooligosaccharide of a degree of polymerization of L to 7 as an excipient. This cellooligosaccharide excipient undoubtedly has water activity which is equal to that of sucrose and is useful as a low-calorie sweetener. According to this patent document, however, since lyophilization is performed when a cellooligosaccharide solution obtained by enzymatic degradation of cellulose is powderized, the cellooligosaccharide powder become light, and the powder fluidity is not necessarily satisfactory. Furthermore, the patent document has no description or suggestion about the crystal shape, powder fluidity, handling properties such as uniform dispersibility of the cellooligosaccharide powder. Therefore, the formulation is essentially different from the cellooligosaccharide powder of the present invention, which highly regulates the crystal shape and physical properties of the powder of cellooligosaccharide; is excellent in powder fluidity; and has improved uniform dispersibility.
Non-patent Documents 3 and 4 describe a cellooligosaccharide powder obtained by enzymatically degrading a sulfite pulp, subjecting the obtained cellooligosaccharide solution to ultrafiltration, purifying with diatom earth and an ion exchange resin then crystallizing with ethanol and isopropyl alcohol. However, crystallization treatment with a poor solvent of the document is intended only to produce cellobiose with higher purity, and there is no description about the crystal shape of cellooligosaccharides. The technique is totally different from that of the present invention for regulating the crystal shape and physical properties of the powder to improve powder fluidity and uniform dispensability of a cellooligosaccharide powder.
Therefore, there have been known no high fluidity cellooligosaccharide powder that has highly regulated saccharide composition and crystal shape of cellooligosaccharides; is excellent in oil retention in addition to powder fluidity and uniform dispersibility; hardly absorbs moisture; and is excellent in handling properties such as a compression moldability, heat resistance, acid resistance, a property of preventing starch retrogradation, and a property of preventing protein modification as that of the present invention, and a method for producing such cellooligosaccharide powder has not been known, too.
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Patent Document 3: JP-A-7-138166
Patent Document 4: JP-A-2006-45186
Patent Document 5: JP-A-11-80781
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